PiT-1/SLC20A1 and PiT-2/SLC20A2 are members of the mammalian type-III Pi transporters encoded by the SLC20 genes. The broad distribution of PiT-1 mRNA and PiT-2 mRNA in mammalian tissues is compatible with housekeeping maintenace of intracellular inorganic phosphate (Pi) homeostasis by transporting Pi from intrastitial fluid for normal cellular functions. Interestingly, mutations of SLC20A2 have been found in patients with idiopathic bilateral basal ganglia and cerebellar calcification (IBGC), also known as Fahr's disease. However, the localization of PiT-1 and PiT-2 in the normal brain has not been clarified yet. The aim of this study was to reveal the distribution of PiT-1 and PiT-2 in the mouse and human brain. Gene expressing analysis showed that PiT-1 and PiT-2 were widely expressed throughout the mouse and human brain, although other Pi transporters encoded by SLC17 and SLC34 genes were hardly detected. Additionally, the region of cerebellum contained a higher level of PiT-1 mRNA and PiT-2 mRNA than the other brain regions. The immonohistochemical studies showed that PiT-1 and PiT-2 was recognized in neuron colocalized with class III β-tubulin, astrocytes identified with GFAP and vascular endothelial cells identified with PECAM-1. Moreover, PiT-1 was distinctly colocalized with PiT-2 in these cells. Our results implicate SLC20 family plays a role in the maintenance of cellular Pi homeostasis especially in neurons, astrocyte and endothelial cells. The findings are compatible with the fact that calcifications are recognized only in the brain. The functional relationship between PiT-1 and PiT-2 remains to be elucidated for the clarification of the pathological mechanism in Fahr's disease.